人参多糖的分级及其免疫活性初探

目的:采用柱分离对人参多糖进行分级,并考察各级分对脾淋巴细胞增殖的刺激作用,找出活性部位.方法:用快流速Q-琼脂糖凝胶( Q-Sepharose FF)色谱将人参多糖进行分级,先用水和0.4 mol·L - 1 NaCl溶液洗脱,将人参多糖分成中性多糖和酸性多糖,再将酸性糖依次用0 ～0.4 mol·L-1NaCl溶液洗脱,得到不同的级分.取小鼠脾脏制备脾淋巴细胞,调整小鼠脾淋巴细胞密度至3.0×106/mL,加样品混合均匀,孵育48 h,考察每个级分不同浓度(25,50,100,200,400 mg·L -1)对脾淋巴细胞增殖的刺激作用.结果:从GP中分级得到GPN和GPA,GPA分级后得到GPA-1,GPA-2,GPA-3和GPA-4.GP,GPA,GPN,GPA-2,GPA-3和GPA-4对脾淋巴细胞增殖都具有促进作用,其最大刺激指数分别为1.073,1.082,1.119,1.101,1.102和1.296.GPA,GPA-2,GPA-3和GPA-4的刺激作用均随着浓度的增加而增强；GPN随着浓度的增加促进作用减弱,在大于100 mg·L-1时呈现抑制作用,且浓度越大抑制作用越强.GPA-1对脾淋巴细胞的增殖具有抑制作用,且浓度越大抑制作用越强.结论:Q-Sepharose FF色谱可将酸性不同的人参多糖逐级分离.人参多糖各级分对脾淋巴细胞增殖有不同程度的促进作用,以GPA-4的刺激作用最强.

Fractionation of Ginseng Polysaccharides and Primary Study of Immune Activity

Objective: Fractionation of ginseng polysaccharides were done by column chromatographic,and study the stimulation of collected fractions on proliferation of spleen lymphocyte to find out the active fractions. Method: Fractionation of ginseng polysaccharides was done by Q-Sepharose FF, separated GP into neutral polysaccharide and acidic polysaccharide with water and 0.4 mol·L^-1 NaCl as eluents,then separated GPA into several fractions with 0-0.4 mol·L^-1 NaCl as eluents. Spleen lymphocytes of mice was isolated and adjusted the spleen lymphocytes concentration to 3.0×10⁶/mL, added sample and incubated for 48 h, the stimulation of each fractions with different concentration(25,50,100,200,400 mg·L^-1)on proliferation of spleen lymphocyte was observed. Result: Obtained GPN and GPA from GP, separate GPA into GPA-1,GPA-2,GPA-3 and GPA-4.GP,GPA,GPN,GPA-2,GPA-3 and GPA-4 had the promoting function on proliferation of spleen lymphocyte,their biggest stimulating indexes were 1.073,1.082,1.119,1.101,1.102 and 1.296. The effects of GPA,GPA-2,GPA-3 and GPA-4 enhanced along with the increasing of concentration;with the increasing of concentration, the promoting action of GPN was reduced,when the concentration was over 100 mg·L^-1, inhibition was found, and with the increasing of concentration,the inhibition became stronger. GPA-1 had suppression inhibitory effects on proliferation of spleen lymphocyte, and with the increasing of concentration, the inhibition became stronger. Conclusion: Different polysaccharide acid were separated from GP with Q-Sepharose FF. Fractionations of ginseng polysaccharides have different promoting function on proliferation of spleen lymphocyte,and GPA-4 had the strongest effect.